• 제목/요약/키워드: Artificial protease

검색결과 19건 처리시간 0.018초

Isolation and Characterization of a Protease-Producing Bacterium, Bacillus amyloliquefaciens P27 from Meju as a Probiotic Starter for Fermented Meat Products

  • Lee, Mi-Sun;Lee, Na-Kyoung;Chang, Kyung-Hoon;Choi, Shin-Yang;Song, Chi-Kwang;Paik, Hyun-Dong
    • 한국축산식품학회지
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    • 제30권5호
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    • pp.804-810
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    • 2010
  • This study was performed to select protease-producing Bacillus sp. as a potential probiotic starter for fermented meat products. In order to isolate protease-producing bacterium from meju, measured the diameter of the clear zone on agar plate (TSA, 1% (w/v) skim milk) and analyzed for intracellular protease activity, then 10 Bacillus-like strains were isolated. Three Bacillus-like strains (P19, P27, and P33) among 10 strains were able to tolerate in acidic condition (TSB, pH 2.5, 2 h incubation). These 3 strains were showed antimicrobial activity against food-borne pathogenic bacteria. These vegetative cells of 3 strains were showed a survival rate of 0.04% to 0.08% under the artificial gastric acidic condition (TSB, pH 2.5 with 1% (w/v) pepsin), but spore-forming cells were 56.29% to 84.77%. Vegetative cells of 3 strains were the least bile-resistant, while spore-forming cells of 3 strains showed higher survival rate more than 76% under artificial bile condition (TSB, 0.1% (w/v) oxgall bile). In these strains, P27 strain was finally selected as a good probiotic strain. P27 strain was tentatively identified as Bacillus amyloliquefaciens by API CHB kit and 16S rDNA sequence analysis. The results of this study suggest that B. amyloliquefaciens P27 can be used as a potential probiotic starter for fermented meat product.

Vibrio vulnificus CYK 279H에서 Collagenolytic Protease 생산을 위한 최적배양조건 (The Optimal Culture Condition for the Collagenolytic Protease Production from Vibrio vulnificus CYK279H)

  • 강성일;김영문;장영부;임동중;공재열
    • KSBB Journal
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    • 제19권4호
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    • pp.295-300
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    • 2004
  • 해양유래미생물에서 gelatin 분해능이 우수한 균주 Vibrio vulnificus CYK 279H를 이용하여 최적 효소활성조건을 검토하였다. 온도와 초기 pH는 $25^{\circ}C$, 7.5에서 높은 것으로 나타났으며, 단당, 이당, 다당류를 첨가하여 탄소원의 영향에서는, 0.3% (w/v) galactose 에서, 유$.$무기 질소원으로는 0.6% (w/v) yeast extract와 4.0% (w/v) gelatin, 0.2% (w/v) (NH$_4$)$_2$SO$_4$에서 효소활성이 가장 높게 나타났다. 염 농도로는 2.0% (w/v) NaCl, 금속이온은 Fe$^{2+}$를 첨가하였을 때 효소활성이 증가되었다. 선정된 최적배양조건에서 Vibrio vulnificus CYK 279H를 배양한 결과, 18 h 배양시 73 Unit/1로 기본배지 20 Unit/1보다 활성이 3.7배 증가한 것으로 확인되었다.

프로테아제와 리파제가 직물의 세척에 미치는 영향 (The Effects of Protease and Lipase on the Detergency of Fabrics)

  • 이정숙;정소화
    • 한국의류산업학회지
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    • 제2권4호
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    • pp.339-345
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    • 2000
  • The effects of protease and/or lipase on the removal of protein soil and oily soil were investigated in this study. Cotton, rayon, nylon, and PET fabrics were soiled by padding of fresh bovine blood and spotting of mixed artificial sebum evenly. The soiled fabrics were aged at $130^{\circ}C$ for 30 minutes. The fabrics were washed by using Terg-O-Tometer at various conditions. Protease and/or lipase were added in the alcohol ethoxylate (AE) detergent solution. The removal efficiency was evaluated by analysis of protein and/or oil on the fabrics before and after washing, respectively. The detergency of protein and/or oil on the fabrics was discussed with enzyme concentration, washing time, washing temperature, pH of washing solution and fiber characteristics. The hydrolysis of protease improved effectively the removal of oil as well as protein by increasing removal of protein-oil mixed soil at the same time. The effect of lipase added detergent solution was slightly shown on the removal of oil and/or protein. The removal of mixed soils from cotton fabrics was very low because of large amount of residual soils caused by the physical characteristics of cotton fiber.

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Overproduction of Streptomyces griseus Protease A and B Induces Morphological Changes in Streptomyces lividans

  • Chi, Won-Jae;Kim, Jung-Mee;Choi, Si-Sun;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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    • pp.1077-1086
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    • 2001
  • The sprA and sprB gene encoding chymotrypsin-like proteases Streptomyces griseus protease A (SGPA) and Streptomyces griseus protease B (SGPB) and the sprT gene that encodes Streptomyces griseus trypsin (SGT) were cloned from Streptomyces griseus ATCC10137 and overexpressed in Streptomyces lividans TK24 as a heterologous host. The chymotrypsin activity of tole culture broth measured with the artificial chromogenic substrate , N-succinyl-ala-ala-pro-phe-p-nitroanilide, was 10, 14 and 14 units/mg in the transformants haboring the sprA, sprB and sprD genes, respectively. The growth of S. lividans reached the maximum cell mass after 4 days of culture, yet SGPA and SGPD production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The trypsin activity of the culture broth measured with the artificial chromogenic substrate , N-${\alpha}$-benzoyl-DL- arginine-p-nitroanilide , was 16 units/mg and SGT production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The introduction of the sprA gene into S, lividans TK24 triggered the biosynthesis of pigmented antibiotics, actinorhodin and undecylprodigiosin, and induced significant morphological changes in the colonies in Benedict, R2YE, and R1R2 media. In addition, the introduction of the sprT gene also induced morphological changes in the colony shape without affecting the antibiotic production, thereby implying that certain proteases would appear to play very important and specific roles in secondary-metabolites formation and morphological differentiation in Streptomyces.

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Pseudoalteromonas donghaensis HJ51의 체외 단백질 분해효소 특성과 생산 조건 (Characterization and optimum production condition of extracellular protease from Pseudoalteromonas donghaensis HJ51)

  • 오지성;최윤수;노동현
    • 미생물학회지
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    • 제51권1호
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    • pp.75-80
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    • 2015
  • 동해에서 분리된 Pseudoalteromonas donghaensis HJ51는 체외분비 단백질 분해효소를 생산하는 신종 미생물로 보고되었다. 체외 단백질 분해효소의 특성과 최적 생산 조건을 결정하기 위해 crude supernatant을 사용하여 단백질 분해효소의 최적 활성 온도와 pH를 조사한 결과 $40^{\circ}C$와 pH 7.5-10.5이었으며, pH 11에서도 88%의 높은 상대적인 효소 활성을 나타내었다. 효소의 금속 요구성을 조사한 결과, $Fe^{3+}$를 10 mM로 첨가하였을 때 최대 효소활성 증가를 보였다. 최대의 효소생산 조건을 탐색한 결과, 기본배지인 PY-ASW (peptone 0.5%, yeast extract 1.0%, artificial seawater)에 탄소원을 첨가하지 않는 것이 가장 높았으며, 질소원으로는 peptone 보다 beef extract, tryptone, casmino acids을 각각 첨가했을 때 활성이 21, 7, 4% 증가하였다. 이러한 결과를 종합해 볼 때 P. donghaensis HJ51이 생산하는 효소는 알칼리성 pH 환경 및 저온환경에서 활성이 필요한 분야에 응용이 가능할 것으로 사료된다.

Artificial Metalloproteases with Broad Substrate Selectivity Constructed on Polystyrene

  • Ko, Eun-Hwa;Suh, Jung-Hun
    • Bulletin of the Korean Chemical Society
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    • 제25권12호
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    • pp.1917-1923
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    • 2004
  • Although the proteolytic activity of the Cu(II) complex of cyclen (Cyc) is greatly enhanced upon attachment to a cross-linked polystyrene (PS), the Cu(II)Cyc-containing PS derivatives reported previously hydrolyzed only a very limited number of proteins. The PS-based artificial metalloproteases can overcome thermal, mechanical, and chemical instabilities of natural proteases, but the narrow substrate selectivity of the artificial metalloproteases limits their industrial application. In the present study, artificial metalloproteases exhibiting broad substrate selectivity were synthesized by attaching Cu(II)Cyc to a PS derivative using linkers with various structures in an attempt to facilitate the interaction of various protein substrates with the PS surface. The new artificial metalloproteases hydrolyzed all of the four protein substrates (albumin, myoglobin, ${\gamma}$-globulin, and lysozyme) examined, manifesting $k_{cat}/K_m$ values of 28-1500 $h_{-1}M_{-1}$ at 50 $^{\circ}C$. The improvement in substrate selectivity is attributed to steric and/or polar interaction between the bound protein and the PS surface as well as the hydrophobicity of the microenvironment of the catalytic centers.

누에인공사료의 사료효율증진에 관한 연구 (Studies on the Efficiency of the Artificial Diet for the Silkworm, Bombyx mori L.)

  • 김주읍
    • 한국잠사곤충학회지
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    • 제28권2호
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    • pp.38-47
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    • 1986
  • 누에 인공동류에 망초, 결명차, 방둠산이 및 동부의 엽분말을 혼입하여 조제한 동료로 누에를 동육한 결과와 그 누에의 체액 중장 견사선등의 전기영동상 및 공식식물의 성분분석내용을 다음과 같이 요약한다. 1. 기본누에인공동료에 망초, 결명차, 방둠산이, 동부의 엽분말을 건물동으로 2-5%정도 혼입한 구에서 섭식과 소화가 조장되었으며 누에의 각종 실용형질도 양호한 경향이었으며 특히 동부와 방둠산이는 첨가효과가 두드러지게 나타났다. 2. 4종의 공식식물엽분말을 2-4종씩 여러 조합으로 혼입한 동료로 누에를 동육한 결과 누에의 섭식과 소화에 대한 각식물성분간의 상승효과는 인정되지 아니하였다. 3. 누에의 채액, 중상 및 견사선의 Esterase Protease Phosphatase의 전기영동상은 처리간에 다소 차이가 인정되었다. 일반적으로 섭식과 소화가 양호한 구에서는 1-2개의 영동대가 더 검출되는 경향이 있었다. 4. 공식식물성분은 상엽에 비해서 전분조지방조단백질무기염류등의 함유율이 현저히 높았으며 섭식에 직접 관여하는 휘발성성분은 찾아내지 못하였다.

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